Quantitative Analysis of hOGG1 Activity Based on Molecular Beacon

doi:10.3969/j.issn.0251-0790.2012.03.010

Abstract

Abstract:

Human 8-oxo-guanine DNA glycosylase 1(hOGG1), which mainly catalyzes the excision of 8-oxoG and cleaves the AP site from DNA to sustain the genome integrity, plays an important role in many biological events including tumor development. However, available methods of hOGG1 assay based on gel electrophoresis are complicate, laborious and discontinuous. Here, we report a new method to analyze hOGG1 activity using molecular beacon containing 8-oxoG coupled with enzyme-linkage reactions, the detection limitation of this method is 0.0125 U/mL. Furthermore, the method was applied for quantification of hOGG1 activity in tumor cells and investigating the effect of metal ions on the reaction. The results demonstrate that the easy, simple and non-radioactive method has the potential to screen suitable inhibitors for hOGG1 and can be applied for high-throughput analysis of hOGG1 activity in tumor samples.

Key words: Molecular beacon, Human 8-oxo-guanine DNA glycosylase1(hOGG1), Real-time monitoring

CLC Number:

O657

TrendMD:

LIU Bin, YANG Xiao-Hai, WANG Ke-Min, TAN Wei-Hong. Quantitative Analysis of hOGG1 Activity Based on Molecular Beacon[J]. Chem. J. Chinese Universities, 2012, 33(03): 486.

References

[1] Achanta G., Huang P.. Cancer Res.[J], 2004, 64(17): 6233-6239
[2] Salem M. M., Shalbaf M., Gibbons N. C., Chavan B., Thornton J. M., Schallreuter K. U.. FASEB J.[J], 2009, 23(11): 3790-3807
[3] Youn C. K., Song P. I., Kim M. H., Kim J. S., Hyun J. W., Choi S. J., Yoon S. P., Chung M. H., Chang I. Y., You H. J.. Mol. Cancer Res.[J], 2007, 5(10): 1083-1098
[4] Zhang H. H., Mizumachi T., Carcel-Trullols J., Li L. W., Naito A., Spencer H. J., Spring P. M., Smoller B. R., Watson A. J., Margison G. P., Higuchi M., Fan C. Y.. Carcino.[J], 2007, 28(8): 1629-1637
[5] Rachek L. I., Thornley N. P., Grishko V. I.. Diabetes[J], 2006, 55(4): 1022-1028
[6] Kondo S., Toyokuni S., Tanaka T., Hiai H., Onodera H., Kasai H., Imamura M.. Clin. Cancer Res.[J], 2000, 6(4): 1394-1400
[7] Gackowski D., Speina E., Zielinska M., Kowalewski J., Rozalski R., Siomek A., Paciorek T., Tudek B., Olinski R.. Cancer Res.[J], 2003, 63(16): 4899-4902
[8] Obtulowicz T., Swoboda M., Speina E., Gackowski D., Rozalski R., Siomek A., Janik J., Janowska B., Ciesla J. M., Jawien A., Banaszkiewicz Z., Guz J., Dziaman T., Szpila A., Olinski R., Tudek B.. Mutagenesis[J], 2010, 25(5): 463-471
[9] Karahalil B., Bohr V., de Souza-pinto N.. Anticancer Res.[J], 2010, 30(12): 4963-4971
[10] Norman D. P., Bruner S. D., Verdine G. L.. J. Am. Chem. Soc.[J], 2001, 123(3): 359-360
[11] LIU Ling-Feng(刘凌凤), TANG Zhi-Wen(唐志文), WANG Ke-Min(王柯敏), LI Jun(李军), GUO Qiu-Ping(郭秋平), MENG Xiang-Xian(孟祥贤), HUANG Shan-Sheng(黄杉生), LI Du(李杜). Chem. J. Chinese Universities(高等学校化学学报)[J], 2003, 24(10): 1761-1764
[12] Tang Z. W., Wang K. M., Tan W. H., Ma C. B., Li J., Liu L. F., Guo Q. P., Meng X. X.. Nucleic Acids Res.[J], 2005, 33(11): e97-1-e97-6
[13] Li J. J., Chu Y. Z., Lee B. Y., Xie X. S.. Nucleic Acids Res.[J], 2008, 36(6): e36-1-e36-17
[14] MENG Xiang-Xian(孟祥贤), YANG Xiao-Hai(羊小海), WANG Ke-Min(王柯敏), GUO Qiu-Ping(郭秋平), LI Jun(李军), TANG Zhi-Wen(唐志文). Chem. J. Chinese Universities(高等学校化学学报)[J], 2009, 30(8): 1538-1542
[15] Ishchenko A. A., Deprez E., Maksimenko A., Brochon J. C., Tauc P., Saparbaev M. K.. Proc. Natl. Acad. Sci.[J], 2006, 103(8): 2564-2569
[16] Matsumoto N., Toga T., Hayashi R., Sugasawa K., Katayanagi K., Ide H., Kuraoka I., Iwai S.. Nucleic Acids Res.[J], 2010, 38(7): e101-1-e101-8
[17] Vidal A. E., Hickson I. D., Boiteux S., Radicella J. P.. Nucleic Acids Res.[J], 2001, 29(6): 1285-1292
[18] Morland I., Luna L., Gustad E., Seeberg E., Bjras M.. DNA Repair[J], 2005, 4(3): 381-387
[19] Adhikari S., Toretsky J. A., Yuan L. S., Roy R.. J. Biol. Chem.[J], 2006, 281(40): 29525-29532
[20] HE Chun-Mei(何春梅), LIU Bin(刘斌), WANG Ke-Min(王柯敏), TANG Hong-Xing(唐红星), HE Li-Fang(何丽芳), LI Hui-Min(李惠敏). J. Chinese Physician(中国医师杂志)[J], 2006, 16(9): 1189-1192
[21] Kameyama K., Huang C. L., Liu D., Masuya D., Nakashima T., Sumitomo S., Takami Y., Kinoshita M., Yokomise H.. Clin. Cancer Res.[J], 2003, 9(13): 4926-4934